Goat TrueBlot®: Anti-Goat IgG HRP
Goat TrueBlot® IP / Western Blot
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Goat TrueBlot®: Anti-Goat IgG HRP

Mouse Monoclonal eB270

24 References
18-8814-31 18-8814-33
50 µL 200 µL
Liquid (sterile filtered) Liquid (sterile filtered)
WB, IP
Goat
Mouse
$129.00 /Per Item
$260.00 /Per Item
50 µL $129.00
200 µL $260.00
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Product Details

Goat TrueBlot®: Anti-Goat IgG HRP - 18-8814-31
Anti-Goat IgG HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting
Mouse
Peroxidase (HRP) ULTRA
Monoclonal
IgG
Immunoprecipitation Kit

Target Details

Goat
Goat TrueBlot® Antibody Peroxidase Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against Anti-Goat Serum. Reactivity is observed against native Goat IgG by both Western blot and ELISA.

Application Details

IP, WB
Goat IgG HRP TrueBlot has been tested in western blot and immunoprecipitation and may also be used for detection in immunoblotting assays that do not employ immunoprecipitation. Goat IgG TrueBlot® is provided as 1000X solution. To conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). If used conservatively at 2.5mls/blot will yield enough reagent for 20 blots. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user. Note that there are three key procedural considerations: 1. Protein A or G beads may be used with the Mouse, Goat and Sheep TrueBlot secondaries, but not with the Rabbit TrueBlot secondary. Use of protein A or G beads with the Rabbit TrueBlot will result in contaminating bands. 2. Immunoprecipitate should be completely reduced. 3. BLOTTO/Milk should be used as the blocking protein for the immunoblot.

Formulation

1 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Proclin is added as an antimicrobial agent.
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol

Shipping & Handling

Wet Ice
Store at -20°C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.
Expiration date is six (6) months from date of receipt.
Goat IgG TrueBlot® is a unique horseradish peroxidase conjugated Anti-Goat IgG monoclonal secondary antibody. Goat IgG TrueBlot® enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Western Blot data with Goat IgG TrueBlot®, simply substitute the conventional HRP Anti-Goat IgG blotting reagent with Goat IgG TrueBlot® and follow the prescribed protocol for sample preparation and immunoblotting. Goat IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of Goat IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Goat IgG TrueBlot® with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/immunoblot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.
(). Inhibition of autocrine HGF maturation overcomes cetuximab resistance in colorectal cancer. Cell Mol Life Sci.
Applications
Other
(). The mechanisms to dispose of misfolded proteins in the endoplasmic reticulum of adipocytes. Nat Commun.
Applications
IP, Co-IP
(). Dominant-Negative Form of SIGIRR: SIGIRRΔE8 Promotes Tumor Growth Through Regulation of Metabolic Pathways. J Interferon Cytokine Res.
Applications
IP, Co-IP; WB, IB, PCA
(). Allosteric potentiation of GABAA receptor single-channel conductance by netrin-1 during neuronal-excitation-induced inhibitory synaptic homeostasis. Cell Rep.
Applications
WB, IB, PCA
(). Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease. Elife.
Applications
WB, IB, PCA
(). Defective NET clearance contributes to sustained FXII activation in COVID-19-associated pulmonary thrombo-inflammation. eBioMedicine
Applications
WB, IB, PCA
(). Angiocrine IGFBP3 spatially coordinates IGF signaling during neonatal cardiac regeneration. bioRxiv Preprint
Applications
WB, IB, PCA
(). Angiocrine IGFBP3 spatially coordinates IGF signaling during neonatal cardiac regeneration. bioRxiv Preprint
Applications
WB, IB, PCA
(). Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins. STAR Protoc.
Applications
WB, IB, PCA
(). Assembly of vascular smooth muscle cells in 3D aggregates provokes cellular quiescence. Exp Cell Res.
Applications
WB, IB, PCA
(). Periostin and tenascin-C interaction promotes angiogenesis in ischemic proliferative retinopathy. Sci Rep.
Applications
IP, Co-IP; WB, IB, PCA
(). Broad-spectrum receptor tyrosine kinase inhibitors overcome de novo and acquired modes of resistance to EGFR-targeted therapies in colorectal cancer. Oncotarget
Applications
WB, IB, PCA
(). Adipose tissue-derived stromal cells are sources of cancer-associated fibroblasts and enhance tumor progression by dense collagen matrix. Int J Cancer.
Applications
WB, IB, PCA
(). Pink1 regulates FKBP5 interaction with AKT/PHLPP and protects neurons from neurotoxin stress induced by MPP. J Neurochem.
Applications
WB, IB, PCA
(). Environmental allergens induce allergic inflammation through proteolytic maturation of IL-33 Nat Immunol
Applications
WB, IB, PCA
(). Selected missense mutations impair frataxin processing in Friedreich ataxia. Annals of Clinical and Translational Neurology
Applications
IP, Co-IP; WB, IB, PCA
(). Improvement of Pharmacokinetic Profile of TRAIL via Trimer-Tag Enhances its Antitumor Activity in vivo. Scientific Reports
Applications
WB, IB, PCA
(). Borrelia burgdorferi BBK32 Inhibits the Classical Pathway by Blocking Activation of the C1 Complement Complex. PLOS Pathogens
Applications
IP, Co-IP
(). The nuclear form of glutathione peroxidase 4 colocalizes and directly interacts with protamines in the nuclear matrix during mouse sperm chromatin assembly. Spermatogenesis.
Applications
WB, IB, PCA
(). High-content analysis of proapoptotic EphA4 dependence receptor functions using small-molecule libraries. J Biolmol Screen
Applications
WB, IB, PCA
(). Host genetics and Chlamydia disease: prediction and validation of disease severity mechanisms. PLoS One.
Applications
WB, IB, PCA
(). Conversion of vascular endothelial cells into multipotent stem-like cells. Nat Med.
Applications
IP, Co-IP
(). PLTP regulates STAT3 and NFκB in differentiated THP1 cells and human monocyte-derived macrophages. Biochim Biophys Acta.
Applications
WB, IB, PCA
(). The p47 GTPases Iigp2 and Irgb10 regulate innate immunity and inflammation to murine Chlamydia psittaci infection. J Immunol.
Applications
WB, IB, PCA

What is Rockland’s TrueBlot® product line?

Rockland’s TrueBlot® product line is designed to solve common experimental problems when performing immunoprecipitation/co-immunoprecipitation experiments and Western blots of immunoprecipitated samples (IP–Western blot). The product line consists of TrueBlot® monoclonal secondary antibodies and TrueBlot® IP beads.

What is the advantage of TrueBlot® secondaries over regular secondary antibodies?

TrueBlot® antibodies are specific to the whole IgG molecule and will not bind heavy or light chains. This is useful for binding target proteins with an expected MW near 25 kDa (light chains) and 50 kDa (heavy chains).

Why does it appear that the TrueBlot® antibody is binding heavy/light chains in my sample?

The sample must be fully reduced to eliminate cross-reactivity with heavy and light chains. Any reactivity to heavy and light chains could be due to incomplete reduction of your sample. Please be sure to optimize reducing conditions for your sample type.

How can I ensure my samples are fully reduced?

Samples can be fully reduced by heating at 90-100°C for 10 minutes in sample buffer containing reducing agent (for example, DTT to a final concentration of 50 mM or add β-Mercaptoethanol to a final concentration of 2% (v/v)).

With which species/isotypes are TrueBlot® secondary antibodies reactive?

TrueBlot® secondary antibodies are reactive with the IgG of their respective species. For example, if using a primary antibody raised in mouse (mouse host), use TrueBlot® anti-mouse Ig secondary antibodies for detection.

Do TrueBlot® secondary antibodies detect IgM?

TrueBlot® secondary antibodies have not been shown to detect IgM.

Do I need to preclear my lysate before the immunoprecipitation step?

Samples that have many other proteins present (such as lysates) may require preclearing to prevent interference in later IP and WB procedures. Recombinant protein samples require pre-clearing more often than serum samples.

How can I enrich/concentrate my lysate for a low expression of a protein of interest?

The immunoprecipitation procedure can be repeated several times to yield a more concentrated protein solution.

What is the recommended lysis buffer for TrueBlot® products?

The optimal lysis buffer will depend on your sample type. See protocol for buffer recommendations. Generally, 1X RIPA is used to lyse samples.

My target protein has the same MW as a heavy/light chain. How can I be sure that the band is the target protein and not a heavy/light chain?

Be sure to include a positive control for the primary antibody in your experiment and check that the sample is fully reduced.

What is the average size of TrueBlot® magnetic/agarose beads?

The beads are roughly 0.5 μm in diameter.

Why can’t I use Protein A or Protein G beads for the IP step in IP–Western blot when using rabbit species?

Using Protein A or Protein G beads with rabbit species results in contaminated bands. For best results when using rabbit species, use Rockland’s TrueBlot® Rabbit Ig IP beads, which are available in either a magnetic or agarose format. Please see individual TrueBlot® Rabbit product pages for additional details.

Can I use Protein A or Protein G beads for the IP step in IP–Western blot when using mouse, goat, or sheep species?

Yes. If you are using Protein A or Protein G beads with mouse, goat, or sheep species, we recommend the following: Determine the compatibility of Protein A or Protein G with your species and subtype, do not use excessive amounts of slurry, and choose the appropriate elution conditions. In some instances, harsher elution conditions can cause stripping of the subunits of Protein A or Protein G from the bead support and result in non-specific bands. For best results, we recommend using TrueBlot® Ig IP beads, which are available in either agarose or magnetic format.

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This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.

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